N-Terminomics/TAILS Profiling of Proteases and Their Substrates in Ulcerative Colitis.

Marilyn H Gordon, Anthonia Anowai, Daniel Young, Nabangshu Das, Rhiannon I Campden, Henna Sekhon, Zoe Myers, Barbara Mainoli, Sameeksha Chopra, Peter S Thuy-Boun, Jayachandran Kizhakkedathu, Gurmeet Bindra, Humberto B Jijon, Steven Heitman, Robin Yates, Dennis W Wolan, Laura E Edgington-Mitchell, Wallace K MacNaughton, Antoine Dufour

Journal: ACS chemical biology 2020;14(11):2471-2483

PMID: 31393699

Full Text on American Chemical Society View on PubMed

Abstract

Dysregulated protease activity is often implicated in the initiation of inflammation and immune cell recruitment in gastrointestinal inflammatory diseases. Using N-terminomics/TAILS (terminal amine isotopic labeling of substrates), we compared proteases, along with their substrates and inhibitors, between colonic mucosal biopsies of healthy patients and those with ulcerative colitis (UC). Among the 1642 N-termini enriched using TAILS, increased endogenous processing of proteins was identified in UC compared to healthy patients. Changes in the reactome pathways for proteins associated with metabolism, adherens junction proteins (E-cadherin, liver-intestinal cadherin, catenin alpha-1, and catenin delta-1), and neutrophil degranulation were identified between the two groups. Increased neutrophil infiltration and distinct proteases observed in ulcerative colitis may result in extensive break down, altered processing, or increased remodeling of adherens junctions and other cellular functions. Analysis of the preferred proteolytic cleavage sites indicated that the majority of proteolytic activity and processing comes from host proteases, but that key microbial proteases may also play a role in maintaining homeostasis. Thus, the identification of distinct proteases and processing of their substrates improves the understanding of dysregulated proteolysis in normal intestinal physiology and ulcerative colitis.

Address: Department of Physiology and Pharmacology , University of Calgary , Calgary , Alberta , Canada T2N 4N1.; Department of Biochemistry and Molecular Biology , University of Calgary , Calgary , Alberta , Canada T2N 4N1.; McCaig Institute for Bone and Joint Health , University of Calgary , Calgary , Alberta , Canada T2N 4N1.; Department of Physiology and Pharmacology , University of Calgary , Calgary , Alberta , Canada T2N 4N1.; McCaig Institute for Bone and Joint Health , University of Calgary , Calgary , Alberta , Canada T2N 4N1.; Department of Biochemistry and Molecular Biology , University of Calgary , Calgary , Alberta , Canada T2N 4N1.; Department of Comparative Biology and Experimental Medicine , University of Calgary , Calgary , Alberta , Canada T2N 4N1.; Departments of Molecular Medicine and Integrative Structural and Computational Biology , The Scripps Research Institute , 10550 North Torrey Pines Road , La Jolla , California 92037 , United States.; Department of Pathology and Laboratory Medicine and Department of Chemistry , University of British Columbia , Vancouver , British Columbia V6T 1Z2 , Canada.; Department of Medicine, Division of Gastroenterology , University of Calgary , Calgary , Alberta , Canada T2N 4N1.; Department of Comparative Biology and Experimental Medicine , University of Calgary , Calgary , Alberta , Canada T2N 4N1.; Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute , The University of Melbourne , Victoria , Parkville , Australia.; Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences , Monash University , Parkville , Victoria , Australia.; Department of Oral and Maxillofacial Surgery , New York University College of Dentistry, Bluestone Center for Clinical Research , New York , New York , United States.; Department of Physiology and Pharmacology , University of Calgary , Calgary , Alberta , Canada T2N 4N1.; Department of Biochemistry and Molecular Biology , University of Calgary , Calgary , Alberta , Canada T2N 4N1.; McCaig Institute for Bone and Joint Health , University of Calgary , Calgary , Alberta , Canada T2N 4N1.

DOI: 10.1021/acschembio.9b00608

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https://doi.org/10.1021/acschembio.9b00608

MeSH Terms: Adult, Aged, Amino Acid Sequence, Binding Sites, Biopsy, Cadherins, Catenins, Chromatography, High Pressure Liquid, Colitis, Ulcerative, Colon, Female, Humans, Isotope Labeling, Male, Mass Spectrometry, Middle Aged, Peptide Hydrolases, Peptides, Protease Inhibitors, Protein Binding, Proteolysis, Proteomics, Signal Transduction